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1. Characterization Of Recombinant Surface Antigen-1 (Rsag-1) Of Toxoplasma Gondii Through Western Blotting Using Mouse Anti-Serum

by Mati Ullah Khan (2008-VA-107) | Dr. Haroon Akbar | Prof. Dr. Khalid Saeed | Dr. Uzma Farid Durrani.

Material type: book Book; Literary form: not fiction Publisher: 2015Dissertation note: Toxoplasmosis is caused by Toxoplasma gondii, which is an obligate intracellular apicomplexan protozoan parasite. The infection is highly prevalent worldwide, and is important both from veterinary and human health concern. In the past recombinant surface antigen-1(rSAG-1) has been shown to be a good candidate for the development of immuno-diagnostic kits as well as Vaccine. Through immuno-blotting, Surface antigen-1 has been identified by sera collected from Toxoplasma gondii infected cats, dogs and humans. Looking at the importance of this health threatening issue, the current study was designed to characterize rSAG-1 through western blotting for development of local diagnostic kit through western blotting using mouse anti-serum. The rSAG-1 was previously expressed under the project of Grand Challenges Canada at Molecular Parasitology laboratory, Department of Parasitology, University of Veterinary and Animal Sciences, Lahore. Recombinant SAG-1 was quantified by using commercial kit based on BCA assay. The 15 μg of rSAG-1 was inoculated subcutaneously (S/C) 3 times with each 2 weeks interval in mice to raise hyper-immune serum. Blood was collected from mice two weeks after the each injection through lateral Retro-Orbital Bleeding (Sharma et al. 2014). Serum was collected by centrifugation. The rSAG-1 was electrophoresed on 12% polyacrylamide gel through SDS-PAGE technique and the protein was transferred to nitrocellulose membrane for western blotting. Anti-serum raised against rSAG-1 was cross-reacted with the rSAG-1 already immobilized on the nitrocellulose membrane. Anti-mice immunoglobulin G conjugated with Alkaline Phosphatase (AP) was used as secondary antibodies for the development of immuno-blot. Immuno-blot revealed a band of 35 KDa. Availability: Items available for loan: UVAS Library [Call number: 2397-T] (1).

2. Molecular Diagnosis Of Anaplasmosis In Buffaloes

by Muhammad Salman (2008-VA-135) | Prof. Dr. Khalid Saeed | Dr. Muhammad Imran Rashid | Dr. Wasim Shehzad.

Material type: book Book; Literary form: not fiction Publisher: 2015Dissertation note: Bovine Anaplasmosis is a tick-borne haemo-rickettsail disease, caused by Anaplasma species transmitted mechanically by flies, biologically by ticks and blood contaminant fomites. It is an economically important tick-borne disease of buffalo in tropical and sub-tropical areas of the world. In current study, we developed and optimized PCR first for detecting Anaplasma at genus level in buffaloes. One hundred (100) blood samples were collected from buffaloes around the Lahore region. The stained thin blood films were examined microscopically and 37% blood samples were found positive for intra-erythrocytic bodies which were then selected for DNA extraction. The DNA was extracted using commercially available kit for eventual use in optimization of PCR for diagnosis of bovine Anaplasmosis. The primers were designed targeting 16S rRNA gene of Anaplasma. For the detection, the PCR product was run in 2% agarose gel stained with ethidium bromide and thirty seven samples showed the amplification band at 179bp. The selected samples were sent for ABI sequencing to Singapore for the accurate detection of the Anaplasma species. The sequencing results were blasted with database of Genbank and we observed homology with Anaplasma phagocytophilum. We found 37% prevalence of Anaplasmosis in buffaloes through PCR. However more studies are required to confirm the species of Anaplasma infecting buffaloes (Bobalus bobalis) by designing species specific primers. Furthermore, additional studies are needed to establish the epidemiology of Anaplasmosis by using molecular tools in different geographical areas of the country for their better control. Availability: Items available for loan: UVAS Library [Call number: 2389-T] (1).

3. Control Of Gastrointestinal Nematodes By Anthelmintics In Buffalo Heifers And Its Effect On Weight Gain

by Tanveer Saroia (2008-VA-062) | Prof. Dr. Khalid saeed | Dr. Nisar Ahmad | Dr. Ali Ahmad Sheikh.

Material type: book Book; Literary form: not fiction Publisher: 2015Dissertation note: Parasitic problem is a major constraint of ruminant production and causes great economic losses to dairy industry by retarded growth, low productivity and increased susceptibility of animals to other infections. The parasitic infections cause productivity losses through reduced feed intake and decreased feed efficiency. Even subclinical or chronic infections are responsible for economic losses in large ruminants. Hypothesis of the study was that administration of anthelmintics will improve weight gain in infected buffalo heifers. One hundred faecal samples of buffalo heifers of age 1-2 years were screened and 58 were found positive for gastrointestinal nematodes. The species of nematodes found prevalent in these animals were Trichostrongylus, Oesophogostomum, Haemonchus, Toxocara, Trichuris and Ostertagia. Thirty buffalo heifers with high EPG were selected and divided into 3 equals groups A, B and C. In this study Group A was a positive control group. Group B was administered Ivermactin (Bomectin) pour on at the dose rate of 1ml / 20 kg body weight. Group C was administered Oxfendazole (Systamex) orally at the dose rate of 1ml/ 10 kg body weight. The effect of these anthelmintics on EPG and weight gain were recorded after every 15 days. This study period was of 90 days (3 months) duration. There was increases in the EPG of positive control group and percent increases was 7.10%, 10.65%, 18.10%, 30.17%, 33.72%, 46.15% at days 15, 30, 45, 60, 75and 90. The percentage decrease in EPG for the group B was 98.22%, 97.04%, 91.12%, 77.51%, 60.94%, 46.74% at days 15, 30, 45, 60, 75 and 90 post treatment and the percentage decrease in EPG for the Group C was 95.80%, 92.81%, 88.62%, 71.85%, 58.68%, 26.34% at days 15, 30, 45, 60, 75 and 90 post treatment. Both anthelmintics cause significant reduction in epg medicated animals as compared to non-medicated animals. There was no difference in the two anthelmintics. The average weight gain per day for Group A (Positive Control) is 0.17 kg per day, for Group B (Ivermectin) is 0.44 kg per day and for Group C (Oxfendazol) was 0.4 kg per day. This shows significant difference between treated groups with non-treated group but there was no difference between the treated groups. It is concluded that ivermectin and oxefendazol can be used for the control of gastrointestinal nematodes. It is recommended that animals may be moved to clean facility after treatment so they are not reinfected. Repeated treatment is suggested to maintain the EPG at low level. Suggestions of the present study are 1. Monitoring of animals for gastrointestinal nematodes by modified McMaster Egg Counting technique. 2. Regular deworming 3. Improved management Availability: Items available for loan: UVAS Library [Call number: 2405-T] (1).

4. Occurrence And Economic Losses From Theileriosis On Commercial Dairy Farm Of Holstein Friesian

by Muhammad Rashid (2014-VA-503) | Dr. Muhammad Imran Rashid | Prof. Dr. Khalid Saeed | Dr. Liaquat Ahmad.

Material type: book Book; Literary form: not fiction Publisher: 2016Dissertation note: Background: Theileriosis is a tick-borne disease and it is transmitted by the bite of ticks. Previous work on disease problems in the study area suggested that Ticks and Tick-Borne Diseases (TTBDs) are the major constraints to cattle production. They cause economic losses to farmers in terms of cattle mortality, loss of body weight, loss of milk production and costs of control of TTBDs by use of acaricides. Theileria is one of the major threat to cattle as it causes anemia, weight loss, decrease production, mortality, treatment cost and cost for the control of theileria. The proper data for losses atributed to theileriosis is still not available in Pakistan. For this purpose a study was carried out in a commercial exotic dairy farm to evaluate losses associated with theileriosis Methodology: The study was done during the period of theileriosis to calculate its economic effect on animal health and production. A total of 150 animals were selected randomly using random number sample formula. The animal tag numbers were compared with random number table, comparing animals were slecteded for study. Thin blood smear was performed for diagnosis haemoparasite, further PCR was performed on those animals that were found +ve for intraerythrocytic bodies. Faecal examination, California mastitis test, teat abnormality and parturition history were recorded for the screening of these factors that decrease milk production. After final grouping, milk production was recorded to identify the effect of theileriosis on production. As theileriosis cause anemia due to destruction of RBC’s. body condition scoring was also performed. Physical examination (lymph node and body temperature) of animals were also performed to evaluate the clinical and subclinical theileriosis. Results: For the evaluation of theileriosis, microscopy was performed on all the animals’ blood samples. Haemoparasites were found in 28.67%. These were further processed by PCR for the CHAPTER 6 SUMMARY Summary 55 detection of theileriosis. Theileria was found in 27.90%. Screening of clinical and subclinical mastitis by Califirnia Mastitis Test and microscopy for gastrointestinal parasite were performed. On faecal examination, there found nematode, cestode and balantidium in 51.72%, 60.92% and 42.53%% respectively. After deworming with Valbazine and curafluke, nematode, cestode (monzia), balantidium and coccidiosis were found in 0%, 39.13, 43.48% and 4.35% respectively. Before grouping clinical and subclinical mastitis were found in 5.38% and 24.62% respectively. After grouping clinical and subclinical mastitis were evaluated by California mastitis test with two weeks interval. At 7th week clinical and subclinical mastitis were 3.85% and 7.69% due to improved management. The decrease in milk production for clinical and subclinical theileriosis was 87 lit./animal and 42.77 lit./animal. Costs for control, treatment and mortality were 0.12%, 0.20% and 13.09% respectively from overall farm expenditure. The prevalence of haemoparasite was 28.67%, while the prevalence of theileriosis was 8%. The new cases of theileriosis were recorded and incidence of theileriosis was found to be 2.25%. Overall losses due to theileriosis was 13.70%. Outcomes: We can conclude from our finding that theileriosis has drastic affect on the profitability of the farms. Then losses can be attributed to decreased milk production and mortality. Medications and control measure for theileriosis have added effect on the losses at exotic animal breed dairy farms. Perspectives: Cost analysis studies need to be done on different dairy farms of cattle of different breeds at different ecological/climatic zones of Pakistan so that investors would know the risks of establishing dairy farms. Availability: Items available for loan: UVAS Library [Call number: 2515-T] (1).

5. Control Of Nematode Infections In Bovines In Different Parity At Calving And Its Effect On Milk Production

by Muhammad Saqib Ali (2008-VA-188) | Prof. Dr. Khalid Saeed | Dr. M. Imran Rashid | Dr. Muhammad Ijaz.

Material type: book Book; Literary form: not fiction Publisher: 2016Dissertation note: Gastrointestinal nematodes are responsible for wide range of health problems, economic losses in bovines and are characterized by reduced milk production, decreased working efficiency and even death. This study was done to evaluate the effect of different anthelmintics on nematode control in different parity cattle at calving and their effect on milk yield. In this study, we screened cattle of 1st and 2nd parity for nematodes. Animals were randomly selected in each group. In first parity animals, we found 23 positive cattle which were divided into 3 different groups while in second parity animals, we found 20 positive cattle which were divided into 3 groups. We used Albendazole (Velbazine) @ 10mg/kg body weight and levamisole (Nilverm) @ 7.5mg/kg bodyweight for treatment. In this study both drugs were found effective in controlling nematodes infections in cattle. Percentage reduction of epg by albendazole was 48.20%, 85.34%, 93.90% and 51.54%, 81.43%, 91.74% on day 7, 14 and 21 in 1st and 2nd parity animals respectively. Percentage reduction of epg by levamisole was 44.45%, 76.92%, 88.03% and 46.60%, 73.78%, 85.43% on day 7, 14 and 21 in 1st and 2nd parity animals respectively. The increase in milk production in current study was 0.39 liter per day and 0.92 liter per day in 1st and 2nd parity cattle respectively. Albendazole increased the milk fat by 0.07 %, while levamisole decreased 0.02 % in 1st parity cattle. In 2nd parity cattle, there was sufficient increase of 0.1% of milk fat by albendazole while levamisole has decreased 0.05 %. We conclude from our studies that anthelmintic treatment of recently calved cattle have significant effect on milk production due to the nematode control. Milk production increased significantly in 2nd parity friesian crossed cattle following anthelmintic treatment as compared to Summary 70 1st parity animals. Levamisole has negative effect on fat concentration in friesian crossed cattle while albendazole treated cattle has positive effect. Albendazole has been found more efficient in reducing EPG of helminths in both parity animals as compared to levamisole treated animals. It is recommended that cattle should be checked for GIT parasites by faecal examination on regular basis so that cattle should be dewormed when needed. Availability: Items available for loan: UVAS Library [Call number: 2527-T] (1).

6. A Comparative Epidemiological Study Of Coccidiosis In Broilers Raised Under Open And Control Sheds

by Shehar Yar Alvi (2007-VA-173) | Prof. Dr. Khalid Saeed | Dr. MUhammad Lateef | Dr. Jawad Nazir.

Material type: book Book; Literary form: not fiction Publisher: 2016Dissertation note: The domesticated fowl (Gallus gallus) is susceptible to seven species of genus Eimeria which are are Eimeria acervulina, Eimeria brunetti, Eimeria maxima, Eimeria mitis, Eimeria necatrix, Eimeria praecox and Eimeria tenella. All of these are capable of causing disease but the clinical picture and pathogenesis may be different according to species, while the pathogenicity ranges from mild to severe. All the species are ubiquitous and cause disease in combination up-to 6 species at the same time on an individual farm so in this sense coccidiosis may be regarded as a disease complex. Now a days subclinical coccidiosis is more frequently affecting the birds as compared to clinical coccidiosis and greatest financial losses are being caused by subclinical coccidiosis in terms of decreased or less weight gain and reduced feed conversion efficiency. . The present study was designed to compile data on the prevalence of coccidiosis in broilers reared under open and controlled sheds situated in and around the Lahore city. Study provided better understanding of the risk factors associated with coccidiosis and their relationship. A questionnaire was designed to record information regarding the management practices, health status of the flock, weight gain. Pooled faecal samples were collected from 50 control sheds and 50 open sheds and were transported to the parasitology laboratory of UVAS. Faecal sample were examined by direct smear to see the coccidial oocysts. Post mortem was conducted to check the presence or absence of the gross lesions associated with coccidiosis. Association between coccidiosis and the risk factors was determined, and the results of open and control sheds were compared. It was assumed that coccidial infections will be higher in the open sheds as compared to environmentally controlled sheds. Open sheds had more prevalence 78% as compared to closed sheds which was reported as 72%. Five major risk factors were studied. Temperature and humidity fluctuation were strong risk factors associated with prevalence of coccidiosis. While litter condition also appeared as an associated risk factor for the prevalence and occurrence coccidiosis in both type of farming systems. Whereas use of medicated feed in open houses appeared as an associated risk factor but in controlled houses use of medicated feed was not associated with the prevalence of coccidiosis. History of previous infections of coccidiosis was also associated risk factor in both type of farming systems. The high prevalence of coccidiosis in open sheds may be due to lack of biosecurity and uncontrollable conditions of temperature and humidity while closed farms have proper biosecurity measures and good husbandry practices. Use of medicated feed and good husbandry practices may be help full to minimize the risk of occurrence of coccidiosis. Further studies are required for better understanding of the disease and associated risk factors. Therefore, the following recommendations are forwarded. • Educating farmers about the importance coccidiosis and its control. • Adaptation of good management practices on farms. • Avoid over-crowding in the house. • Alternative remedies need to be developed and evaluated to prevent and control coccidiosis. Availability: Items available for loan: UVAS Library [Call number: 2691-T] (1).

7. Prevalence And In Vitro Acaricidal Activity Of Nicotiana Tobacum Extract(S) Against Ticks(S) Of Cattle In District Loralai (Balochistan)

by Najeeb Ullah (2008-VA-203) | Dr. Muhammad Lateef | Dr. Saad Ullah Jan | Prof. Dr. Khalid Saeed | Prof. Dr. Aneela Zameeer Durrani.

Material type: book Book; Literary form: not fiction Publisher: 2016Dissertation note: In this study prevalence and acaricidal activity of Nicotiana tobacum plant leaves extracts of chloroform and methanol based was evaluated. 6.1 Prevalence study In this study total 670 cattles were examined for tick infestation in cattle of different breed, age and sex in district Loralai (Balochistan). Overall prevalence of tick infestation of cattle recorded was 21.49%. Breed wise prevalence was 26.15, 12.80 and 22% in Friesian, Sahiwal and non-descriptive breed of cattle respectively. Age wise prevalence was 27.90, 26.88 and 19.34% in <1 year, 2 year and >2 years of cattle respectively. Higher sex related prevalence was noticed in female cattle (21.98%) as low found with male cattle (16.92%). 6.2 Acarididal effects of tobacco (Nicotiana tobacum) plant The plant leaves of Nicotiana tobacum were dried for 8 to 10 days. The leaves were grinded mechanically into powder form and extract was prepared in Soxhlets apparatus. Extract was further dried in rotatory evaporator and hot air oven to remove left over moisture to obtain solid extract. The dry powder was stored in refrigerator at 4 °C to protect it from any fungal contamination. The powder extracts was used to make different concentrations of 12.5mg/ml, 25mg/ml and 50mg/ml in distilled water. The ticks collected from study area was weighed and dipped in to the formulated solution for interval of 5 mints. After immersion ticks were incubated at 30 °C temperature and 80% relative humidity. After the oviposition period (18 days), the eggs were collected and weighed for each group. The comparison of all the groups was observed in terms of egg laying index and percentage inhibition of egg laying. 200 eggs (Approximately 10mg) were studied for egg hatchability of different groups at different concentration of chloroform and methanol extracts, This study has successfully achieved main objectives to determine the acaricidal effects of Nicotina tobacum extracts on ticks of cattle. Egg laying index: values of egg laying index at concentration of 12.5mg/ml, 25mg/ml and 50mg/ml for chloroform and methanol extracts were as follows, for chloroform extracts these were 0.4782800±0.02789077, 0.4388300±.05119868 and 0.3963600±0.03380405 and for methanol extracts these were 0.4991200±0.00948646, 0.4614300±0.03917896 and 0.4205800±0.04183098 respectively. While for control group it was calculated 0.5331200±0.02757486 for all concentrations of Nicotiana tobacum extracts. This decline in egg laying index showed by ticks of chloroform and methanol extracts was significantly different (P<0.05) from control group. Percentage inhibition of egg laying: For chloroform extract calculated value was 10.048, 17.378 and 25.143% at concentration of 12.5mg/ml, 25mg/ml and 50mg/ml respectively, and for methanol extracts the value was 6.367, 13.152 and 20.827% at concentration of 12.5mg/ml, 25mg/ml and 50mg/ml respectively. Egg hatchability: Hatchability of ticks eggs of chloroform extract of Nicotiana tobacum plant were recorded 67.5, 43.5 and 17% at concentration of 12.5mg/ml, 25mg/ml, and 50mg/ml respectively. Moreover, hatchability of ticks eggs of methanol extract of Nicotiana tobacum plant were recorded 77.5, 47.5 and 23% at concentration of 12.5mg/ml, 25mg/ml, and 50mg/ml respectively and for control group it was recorded 100% as treated with distilled water. Availability: Items available for loan: UVAS Library [Call number: 2687-T] (1).

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